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Dynamic Adhesion Assay for the Functional Analysis of Anti-adhesion Therapies in Inflammatory Bowel Disease
Published on: September 20, 2018
Mapping specific adhesive interactions on living human intestinal epithelial cells with atomic force microscopy
A Patrick Gunning1, Stephen Chambers, Carmen Pin
1Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK. patrick.gunning@bbsrc.ac.uk
Summary
Researchers developed a new method using atomic force microscopy to measure molecular interactions on gut cells. This technique maps receptor locations and binding forces, aiding drug delivery and understanding gut health.
Area of Science:
- Biophysics
- Cell Biology
- Gastroenterology
Background:
- Understanding molecular-receptor interactions on gut epithelium is crucial for food bioactivity, drug efficacy, and pathogen defense.
- Current methods for probing these specific interactions on living gut cells are limited.
- Living human colorectal cancer cells serve as valuable in vitro models for gut epithelium research.
Purpose of the Study:
- To develop and validate a novel methodology for measuring specific molecular-receptor interactions on living gut epithelial cells.
- To map the location and distribution of receptors on the cell surface.
- To provide a tool for advancing targeted drug delivery systems and understanding gut health.
Main Methods:
- Utilized Atomic Force Microscopy (AFM) to measure single-molecule ligand-receptor interactions.
- Employed functionalized silica beads on AFM tips to probe binding events.
- Applied the method to living Caco-2 human intestinal epithelial cells using Wheat Germ Agglutinin (WGA) as a ligand.
Main Results:
- Successfully measured the modal detachment force of WGA binding to Caco-2 cells at 125 pN, indicative of single-molecule interactions.
- Mapped the spatial distribution of binding sites on the cell surface.
- Revealed heterogeneity in receptor distribution within and between cells in a monolayer.
Conclusions:
- The developed AFM-based methodology enables precise measurement of molecular-receptor interactions on living gut epithelial models.
- This technique offers insights into receptor localization and binding forces, crucial for targeted therapies.
- The findings contribute to advancing our understanding of gut epithelium function and developing novel therapeutic strategies.
